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Kit and method for detecting furacilin metabolin

A technology of nitrofurazone and metabolites, which is applied in the field of detection of nitrofurazone metabolite residues in animal tissues, can solve the problems of complex processing, high degree of instrumentation, large detection sample volume, etc., and achieve simple operation, high sensitivity, and simple storage Effect

Active Publication Date: 2013-10-23
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most common methods used to detect nitrofuran metabolites are high performance liquid chromatography (HPLC), liquid chromatography-mass chromatography (LC-MS / MS), etc. These methods have high sensitivity, accurate results, good repeatability, and false There are few positives, but the sample pretreatment process is complicated, and the instrumentation is high and expensive. Compared with it, the immunological method has the characteristics of high sensitivity, simple sample pretreatment, short detection time, and large detection sample volume, etc., which is suitable for on-site monitoring. and screening of a large number of samples

Method used

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  • Kit and method for detecting furacilin metabolin
  • Kit and method for detecting furacilin metabolin
  • Kit and method for detecting furacilin metabolin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The experimental methods used in the following examples are conventional methods unless otherwise specified. Embodiment 1 detects the composition of the kit of nitrofurazone metabolite

[0040] 1. Test paper (called test strip) ( figure 1 )

[0041] The test paper is composed of a bottom plate, a sample absorption pad, a reaction film, a water absorption pad, and a protective film;

[0042] The sample absorbent pad 1, the reaction film 2, the water absorbent pad 3 and the protective film 7 are pasted on the bottom plate 6 in sequence, the end of the sample absorbent pad is connected with the reaction film, the end of the reaction film is connected with the water absorbent pad, and the sample absorbent pad The beginning is aligned with the beginning of the bottom plate, and the end of the absorbent pad is aligned with the end of the bottom plate;

[0043] The sample absorption pad end of the test paper is pasted with a protective film, and the protective film 7 cover...

Embodiment 3

[0098] The detection of nitrofurazone metabolite in embodiment 3 animal tissue

[0099] 1. Sample pretreatment

[0100]Weigh 5.0g ± 0.05g homogeneous sample into a 50ml polystyrene centrifuge tube, add 5ml 10% trichloroacetic acid, and then add 100μl derivatization reagent (add to the reagent bottle containing 151mg 2-nitrobenzaldehyde Dissolve 10ml of methanol and mix well), shake fully for 3min; incubate in a 60°C incubator for 1.5h; take out and add 1ml 0.5mol / L dipotassium hydrogen phosphate solution, 1.5ml 2mol / L sodium hydroxide solution and 10ml ethyl acetate, Oscillate for 10s, shake lightly for 8-10 times for samples with high fat content to prevent emulsification; centrifuge at room temperature (20-25°C) for 5 minutes above 3000g; put 8ml of ethyl acetate into a 10ml dry glass test tube, place at 50~ Blow dry in a water bath at 60°C under nitrogen flow; add 1ml of n-hexane, vortex for 30s, add 0.8ml of sample complex solution (0.2mol / L phosphate buffer), vortex for ...

Embodiment 4

[0105] The determination of embodiment 4 kit technical parameter

[0106] 1. Sensitivity test

[0107] The nitrofurazone metabolite standard (purchased from Sigma) was diluted to 0.5, 1.0, 2.0 μg / L; the diluent used was pH7.2, 0.2mol / L phosphate buffer.

[0108] Tested with the kit, the results are: when the standard concentration of nitrofurazone metabolites is 0.5 μg / L, two red lines visible to the naked eye appear on the test strip, which is negative; the concentrations of standard nitrofurazone metabolites are 1.0 and 2.0 μg / L, the quality control area of ​​the test strip develops color, but the detection area does not develop color, which is positive, indicating that the sensitivity of this kit to detect nitrofurazone metabolites is 1.0 μg / L.

[0109] 2. False positive rate, false negative rate test

[0110] Take 20 pork, chicken, fish, and shrimp positive samples with known nitrofurazone metabolite content greater than 1.0 μg / kg and 20 pork, chicken, fish, and shrimp ...

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Abstract

The invention discloses a kit and method for detecting a furacilin metabolin. The kit comprises a microporous reagent and test paper, wherein a specific antibody of the furacilin metabolin, which is marked by collaurum, is freeze-dried in the microporous reagent; the specific antibody of the furacilin metabolin can be taken as a monoclonal antibody or a polyclonal antibody of the furacilin metabolin; the test paper is formed by sequentially connecting a sample absorbing pad, a reaction membrane, a water absorbing pad, a protective membrane and a bottom plate, wherein a detecting area and a quality control area are formed on the reaction membrane; a hapten-carrier protein conjugate of the furacilin metabolin is wrapped in the detecting area; and an antibody is wrapped in the quality control area. The kit for detecting the furacilin metabolin is simple, rapid, visual and accurate in method, wide in application range and low in cost, thereby being easy to popularize.

Description

technical field [0001] The invention relates to a kit and method for detecting nitrofurazone metabolites, in particular to a colloidal gold test strip for detecting nitrofurazone metabolites in a sample, which is particularly suitable for detection of nitrofurazone metabolite residues in animal tissues. Background technique [0002] Nitrofurans are often used as broad-spectrum antibiotics to prevent and treat gastrointestinal diseases in pigs, cattle, poultry and bees caused by Salmonella and Escherichia. However, in the course of long-term experimental research, it was found that both nitrofuran drugs and metabolites can cause cancer and gene mutations in experimental animals, and such drugs are prohibited from being used in treatment and feed. [0003] Since the prototype drug of nitrofuran is rapidly metabolized in the body, it cannot be detected, but its metabolites are quite stable due to the combination with proteins, so when analyzing the residues of such drugs, it is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558
Inventor 何方洋罗晓琴陶光灿吴鹏陈炜玲张荃冯才茂杨秀贤刘玉梅
Owner BEIJING KWINBON BIOTECH
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